Chien-Min Chen1,2, Shu-Ching Hsieh3, Chia-Liang Lin4, Yu-Syun Lin4, Jen-Pi Tsai5,6, Yi-Hsien Hsieh4,7,8. 1. Division of Neurosurgery, Department of Surgery, Changhua Christian Hospital, Changhua, Taiwan. 2. School of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. 3. School of Medical Laboratory and Biotechnology, Chung Shan Medical University, Taichung, Taiwan. 4. Institute of Biochemistry, Microbiology and Immunology, Chung Shan Medical University, Taichung, Taiwan. 5. School of Medicine, Tzu Chi University, Hualien, Taiwan. 6. Division of Nephrology, Department of Internal Medicine, Dalin Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Chiayi, Taiwan. 7. Department of Biochemistry, School of Medicine, Chung Shan Medical University, Taichung, Taiwan. 8. Clinical laboratory, Chung Shan Medical University Hospital, Taichung, Taiwan.
Abstract
BACKGROUND/AIMS: α-mangostin has anti-carcinogenic effects against several cancers. We investigated the molecular mechanism of this compound on the metastasis of human renal carcinoma cells. METHODS: Cell viability was measured using the MTT assay, and cell cycle distribution using flow cytometry. A Matrigel-based assay was used to measure in vitro cell migration and invasion. MAPK-related proteins and matrix metalloproteinase (MMP)-9 and MMP-2 expression were measured by western blotting, and MMP2/-9 activities were determined by gelatin zymography. RT-qPCR and a luciferase assay were used to examine the transcriptional activity of MMP-9. RESULTS: α-mangostin inhibited the migration and invasion of RCC cells in a dose-dependent manner, but had no evident cytotoxic effects. Treatment of 786-O cells with α-mangostin inhibited activation of MEK and ERK. Treatment with a specific MEK inhibitor (U0126) enhanced the inhibitory effects of α-mangostin on cell migration and invasion, and the phosphorylation of ERK and MEK. Moreover, α-mangostin inhibited the expression of the MMP-9 mRNA levels as well as the activity of MMP-9 promoter, and these suppressive effects were further enhanced by U0126. CONCLUSIONS: Our results suggest that α-mangostin suppresses cell migration and invasion via MEK/ERK/MMP9 pathway, and might be a promising anti-metastatic agent against human renal cell carcinoma.
BACKGROUND/AIMS: α-mangostin has anti-carcinogenic effects against several cancers. We investigated the molecular mechanism of this compound on the metastasis of human renal carcinoma cells. METHODS: Cell viability was measured using the MTT assay, and cell cycle distribution using flow cytometry. A Matrigel-based assay was used to measure in vitro cell migration and invasion. MAPK-related proteins and matrix metalloproteinase (MMP)-9 and MMP-2 expression were measured by western blotting, and MMP2/-9 activities were determined by gelatin zymography. RT-qPCR and a luciferase assay were used to examine the transcriptional activity of MMP-9. RESULTS: α-mangostin inhibited the migration and invasion of RCC cells in a dose-dependent manner, but had no evident cytotoxic effects. Treatment of 786-O cells with α-mangostin inhibited activation of MEK and ERK. Treatment with a specific MEK inhibitor (U0126) enhanced the inhibitory effects of α-mangostin on cell migration and invasion, and the phosphorylation of ERK and MEK. Moreover, α-mangostin inhibited the expression of the MMP-9 mRNA levels as well as the activity of MMP-9 promoter, and these suppressive effects were further enhanced by U0126. CONCLUSIONS: Our results suggest that α-mangostin suppresses cell migration and invasion via MEK/ERK/MMP9 pathway, and might be a promising anti-metastatic agent against humanrenal cell carcinoma.
Authors: Min Tao; Yingfeng Shi; Lunxian Tang; Yi Wang; Lu Fang; Wei Jiang; Tao Lin; Andong Qiu; Shougang Zhuang; Na Liu Journal: Am J Physiol Renal Physiol Date: 2019-01-16